The expression of a selected set of miRNAs, which exhibited altered expression in ERG-positive CaP tumor tissues compared to ERG-negative CaP tumors is further validated in two different CaP cell lines, LnTE3 and VCaP cells. LnTE3 cells (LNCaP-lentivirus TMPRESS2:ERG3, inducible) were treated with doxycycline (1μg/ml) for 72 hrs to induce ERG expression: a. protein analyzed by immunoblot and b. TMPRESS2-ERG mRNA analyzed by qPCR. c. The expression of miR-125-5p, miR-520g, miR-874, miR-449a and miR-660 was analyzed by Taqman miR-specific assays in LnTE3 cells. VCaP cells were treated with siERG or control siRNA, both d. ERG protein and e. mRNA were analyzed by immunoblot and qPCR, respectively. f. The expression of miR-125-5p, miR-520g, miR-874, miR-449a and miR-660 was similarly analyzed by miR-specific Taqman assays in VCaP cells. For all miR-specific assays RNU48 was used as an endogenous control. The data reflect averages of at least three independent experiments (* indicates p<0.05).